SEASONAL FLUCTUATIONS IN TISSUE NITROGEN,PHOSPHORUS, AND N:P FOR FIVE MACROALGAL SPECIES COMMON TO THE PACIFIC NORTHWEST COAST1

Variations in tissue nitrogen (N) and phosphorus (P) were examined over a complete seasonal cycle for five macroalgae common in Oregon coastal water. Tissue N ranged from 2.0 to 5.5% dry weight (dry wt) in leafy macroalgae (Enteromorpha intestinalis (Linnaeus) Link. Ulva fenestrata Postels et Ruprecht, and Porphyra sp.) and from 0.9 to 2.6% drt wt in branched macroalgae (Codium fragile (Suringar) Hariot and Pelvetiopsis limitata Setchell). Tissue P ranged from 0.32 to 0.86% dry wt in leafy macroalgae and from 0.27 to 0.50% dry wt in branched macroalgae, Ulva fenestrata, C. fragile, and P. limitata appeared to be N limited during part of the year based on tissue N levels. Variations in N:P showed a more distinct seasonal pattern than either tissue N and tissue P. All macroalgae examined appeared to be N limited at least part of the year based on N:P composition, and P limitation occurred for all macroalgae examined except C. fragile. Our results suggest that tissue N:P ratio for macroalgae may be a good index for evaluating in situ nutrient status.     

GLYCOCONJUGATE ORGANIZATION OF ENTEROMORPHA (=ULVA) FLEXUOSA AND ULVA FASCIATA (CHLOROPHYTA) ZOOSPORES1

Ecologically successful algae that colonize natural and artificial substrates in the marine environment have distinct strategies for opportunistic dispersal and settlement. The objective of this research was to visualize molecular architecture of zoospores from Enteromorpha (=Ulva) flexuosa (Wulfen) J. Agardh and Ulva fasciata Delile that coexist but alternate in dominance on an intertidal bench. Multiple fluorescent lectins were used to stabilize and probe for diverse zoospore glycoconjugates (GC) that could be involved in cell and substrate interactions. Results from epifluorescence microscopy showed distinct cellular and extracellular polymeric substance (EPS) domains of GC relative to settlement morphologies. Glycoconjugates were similar for both species with (1) α‐d mannose and/or glucose moieties localized on flagella, the anterior domes and anterior regions, the plasma membranes, and EPS; (2) α‐fucose localized on flagella and anterior regions; (3) N or α,ß‐N acetylglucosamine localized on flagella, the anterior regions, and EPS; and (4) varied N‐acetylgalactosamine and/or galactose moieties localized on each domain for both species excluding the plasma membranes. Some differences in lectin binding were observed for each species at the flagella, the anterior domes, and the plasma membranes. Glycoconjugate distributions shifted with morphological changes that followed initial adhesion. TEM of E. flexuosa zoospore stages following carbohydrate‐stabilizing fixations and gold‐conjugated lectin probes resolved GC with α‐d mannose and/or glucose, and/or N‐acetylglucosamine at the plasma membrane, ER and diverse vesicles of the anterior pole, EPS, and discontinuous regions or knobs associated with flagellar surfaces. The distinct distribution and diversity of zoospore GC may be central to recognition and attachment on diverse substrata by these algae.     

INHIBITION OF GROWTH OF ULVA EXPANSA (CHLOROPHYTA) BY ULTRAVIOLET-B RADIATION1

We examined the effect of ultraviolet-B radiation (UV-B, 290–320 nm) on the growth rate of the intertidal marine alga Ulva expansa (Setch.) S. & G. (Chlorophyta). Segments of thallus collected from a natural population were grown in outdoor seawater tanks. Combinations of UV-B-opaque screens, UV-B-transparent screens, and UV-B lamps were used to investigate the effects of solar UV-B and solar plus supplemental UV-B on the growth of these segments. Growth was measured by changes in segment surface area, damp weight, and dry weight. Growth rates of segments were inhibited under both solar UV-B and solar plus supplemental UV-B treatments. Growth rates were also inhibited by high levels of photosynthetically active radiation, independent of UV-B fluence. These results indicate that increases in UV-B resulting from further ozone depletion will have a negative impact on the growth of this alga.     

EFFECTS OF LIGHT ON GROWTH,RuBPCase ACTIVITY AND CHEMICAL COMPOSITION OF ULVA SPECIES (CHLOROPHYTA)1

The effects of light on growth, RuBPCase activity, and chemical composition of Ulva curvata (Kütz.) De Toni and U.Lactuca L. were examined at a range of temperatures and N-supply levels. Groeth of Ulva speices becomes more light-dependent with increasing temperature and N. The effect of light on RuBPcase is N-dependent, with a positive correlation under N-sufficient and a negative correlation under N-limited conditions. Light effects on pigment levels and ratios may be independent of effects on growth rate. These interactions uncouple growth rate from RuBPCase and pigments, and thus from tissue%N. The limits of variability of the growth-%N relationship can be described by a parabola. Under relative light or temperature-limitation, %N is negatively, growth increase with increasing %N. Tight coupling of seaweed wrowth and chemical composition may therefor be relatively rare in natural waters where growth can be simultaneously limited by light, temperature, and N.     

PHOTOACCLIMATION AND GROWTH RATE RESPONSES OF ULVA ROTUNDATA (CHLOROPHYTA) TO INTRADAY VARIATIONS IN GROWTH IRRADIANCE1

A vegetative clone of the chlorophyte macroalga Ulva rotundata was maintained in an outdoor continuous flow system under nutrient sufficient conditions and various light regimes. Step changes between 9 and 100% incident irradiance (I^o) were employed to simulate cloud passage. Temporary (1–4 h) midday (I^o)perturbations evoked net changes in growth rate (μ) and chlorophyll (chl) content. Under I_o alternating at various periodicities from 15 min to 7 h, net μ was the average of the μ under steady state 9 and 100% I^o, regardless of periodicity. However, the μ in alternating light was considerably less than μ under steady state 55% I^o(? 9%+ 100%/2), as expected based on the nonlinear shape of the μ-I relationship. Unlike μ, chl content depended primarily on the total daily irradiance, probably clue to the slower response of chl compared to photosynthetic rate. On time scales ≥ one day, chl was linearly correlated with light-regulated daily μ under both steady state and intraday fluctuating irradiance, consistent with photosynthetic feedback regulation of chl concentration.     

GROWTH OF ULVA FENESTRATA (CHLOROPHYTA) IN MICROCOSMS RICH IN ZOSTERA MARINA (ANTHOPHYTA) DETRITUS1,2

In laboratory experiments discs cut from thalli of Ulva fenestrata Postels & Ruprecht grew in diameter and biomass (dry weight and ash-free dry weight) more when dead leaves of Zostera marina L. were present than when absent. A maximum increase in dry weight of 500% in 14 days occurred with Zostera present compared with only a 200% increase with no detritus. When NO^?₃ and PO^3-₄ were added the weight of an Ulva disc increased by over 800% when Zostera was present but by only 400% in controls. In general penicillin G (475 units/ml) caused a reduction in algal growth in cultures containing detritus. It is concluded that bacteria on the detritus may compete with algae for nutrients but by releasing unknown substances the bacteria promote the growth of Ulva.     

MOLECULAR AND MORPHOLOGICAL ANALYSIS OF ENTEROMORPHA INTESTINALIS AND E. COMPRESSA (CHLOROPHYTA) IN THE BRITISH ISLES

The very common green seaweeds Enteromorpha intestinalis (L.) Nees and E. compressa (L.) Nees are important fouling organisms and have commonly been used as indicators of eutrophication, but their taxonomic status is problematic. The genus presents extreme difficulties because there is wide intraspecific variation in morphology, but morphological differences between species are small and difficult to detect. In this study, molecular data were used in parallel with morphological characters to resolve the taxonomic problems. Phylogenetic analysis of sequences of the internal transcribed spacers ITS1 and ITS2 and the 5.8S gene distinguished two groups of samples, which were identified by morphological characters as E. compressa (branched) and E. intestinalis (normally unbranched). There was a low level of sequence divergence within each group of samples, but divergence between groups was as great as that between either of the two species and the outgroup E. prolifera. Clades representing E. compressa and E. intestinalis were also found in analyses of an independent molecular data set, chloroplast DNA restriction fragment length polymorphisms (RFLPs). Enteromorpha intestinalis and E. compressa represent two distinct, genetically divergent species. Reinterpretation of published studies shows that these species are reproductively isolated. However, E. compressa and E. intestinalis are sometimes very difficult to distinguish from each other and could be regarded as cryptic species. The presence or absence of branching was the most useful character distinguishing these two species, but there was an element of ambiguity because low salinity or salinity shock can induce branching in E. intestinalis. If environmental factors such as salinity are taken into account, branching can be used to identify the great majority of thalli correctly. This study therefore provides a basis for identifying the two most important marine fouling macroalgae and for their use in environmental monitoring and experimentation. Typification of these two Linnaean species showed that current usage of the names accords with the lectotype and protologue of both species. Samples that resembled E. usneoides did not form a clade in any of the trees, and constraining the data to support the monophyly of this group incurred a penalty. Enteromorpha usneoides appears to be an ecotype of E. compressa.     

CHANGES IN NITROGEN POOLS IN ULVA FENESTRATA (CHLOROPHYTA) AND GRACILARIA PACIFICA (RHODOPHYTA) UNDER NITRATE AND AMMONIUM ENRICHMENT

The accumulation of nitrogen in different cellular pools by the macroalgae Ulva fenestrata (Postels and Ruprecht) (Chlorophyta) and Gracilaria pacifica (Abbott) (Rhodophyta) was studied in a laboratory experiment. After 8 or 9 days of nitrogen enrichment, nitrate, ammonium, free amino acid (FAA), protein, chlorophyll (chl), phycoerythrin (PE), and insoluble nitrogen pools were extracted and analyzed, and their relative contribution to total nitrogen (TN) was assessed. In U. fenestrata, the nitrate and ammonium enrichments resulted in a significant increase of TN from 2.41% dry weight (dw) to 4.19% and 4.71% dw, respectively. All the extracted N pools increased significantly. In G. pacifica, TN increased more under ammonium enrichment than under nitrate enrichment. In both macroalgae, proteins and FAA were the most important N storage pools. Protein-N ranged from 700 to 2300 μmol N·g dw⁻¹ (43%–66% of TN) and contributed the most to TN increase (41%–89%). The FAA pool was always larger in G. pacifica than in U. fenestrata. In both species, the FAA pool accounted for 4%–17% of TN (70–600 μmol N·g dw⁻¹). In U. fenestrata, nitrate can represent a temporary storage pool: it accumulated up to 200 μmol N·g dw⁻¹ (7% of TN) and contributed more than FAA to overall increase in cellular nitrogen. In contrast, G. pacifica had a small nitrate pool. The PE pool in G. pacifica increased with TN but was never more than 9% of total protein-N or 6% of TN, and it was less important than FAA as a storage pool. All TN was recovered in the extracted and insoluble N pools at the end of the experiment in U. fenestrata. In G. pacifica, the extracted and insoluble N pools accounted on average for 83%–90% of TN.     

THE EFFECTS OF PHOSPHITE ON PHOSPHATE STARVATION RESPONSES OF ULVA LACTUCA (ULVALES,CHLOROPHYTA)1

Effects of phosphite (Phi) on phosphate (Pi) starvation responses were determined in Ulva lactuca L. by incubation in Pi‐limited (1 μM NaH₂PO₄) or Pi‐sufficient (100 μM NaH₂PO₄) seawater containing 0–3 mM Phi. Exposure to 1 μM NaH₂PO₄ decreased the growth rate and the content of free Pi and esterified‐P but increased the activities of extracellular alkaline phosphatase (EC 3.1.2.1) and intracellular acid phosphatase (ACP; EC 3.1.2.2); two ACP isozymes observed by activity staining on isoelectric focussing (IEF) gel were induced. The K_m value of Pi uptake rate was decreased by incubation with 1 μM NaH₂PO₄ and the decrease in K_m value was inhibited by 2 mM Phi, reflecting the operation of a high‐affinity Pi uptake system at low Pi concentrations. In the presence of Phi, the growth rate of Pi‐sufficient and Pi‐starved thalli decreased as Phi concentrations were increased from 0 to 2 mM. As Phi concentrations were increased from 0 to 2 mM, the free Pi contents in both Pi‐sufficient and Pi‐starved thalli decreased, but the esterified‐P contents in Pi‐starved thalli increased, whereas those in Pi‐sufficient thalli increased at 1 mM Phi and decreased at 2 mM Phi. Cell wall localized AP activity in both Pi‐sufficient and Pi‐starved thalli decreased as Phi concentrations were increased from 0 to 2 mM. Intracellular ACP activity in Pi‐starved thalli decreased as Phi concentrations were increased from 0 to 2 mM but was not affected in Pi‐sufficient thalli. The induction of ACP isozyme activity and high‐affinity Pi uptake system in Pi‐starved thalli was inhibited by Phi. The present investigation shows that Phi interrupts the sensing mechanisms of U. lactuca to Pi‐limiting conditions.     

UNCOUPLING OF VARIOUS MEASURES OF GROWTH IN ULVA ROTUXDATA(CHLOROPHYTA) FOLLOWING A LARGE DECREASE IN IRRADIANCE1

A vegetative clone of the chlorophyte macroalga Ulva rotundata Blid. was maintained in an outdoor continuous flow system and subjected to a large decrease in irradiance. Specific growth rates based on changes in fresh (μ_FW) and dry weight (μ_DW) and surface area (μ_SA) were determined using precut disks over the 24 h following a post-sunset transfer from full sunlight (100% I₀) to 9% I₀ All three measures of growth rate were approximately equivalent for untransferred control plants at either limiting (9%) or saturating (100%)I₀. Transferred disks exhibited μ_FW and μ_SA which were slightly lower than 100%I₀ controls and much higher than 9% I₀ controls; μ_DW was nearly identical for transferred disks and 9% I₀ controls. Cell size was unchanged following transfer, indicating that surface area changes reflected a proportional increase in cell number. Cell division therefore continued at a high rate for one day following transfer of U. rotundata to irradiances which are subsaturating for photosynthesis (indicated by μ_Dw). Starch reserves were largely depleted, and the C/N ratio decreased during this period.     

EFFECTS OF TEMPERATURE,NITROGEN SUPPLY,AND TISSUE NITROGEN ON AMMONIUM UPTAKE RATES OF THE CHLOROPHYTE SEAWEEDS ULVA CURVATA AND CODIUM DECORTICATUM1

Nitrogen uptake rates of Ulva curvata (Kütz.) de Toni (Ulvales) and Codium decorticatum (Woodw.) Howe (Caulerpales) grown under several N addition regimes were determined by perturbation and continuous mode techniques, and as N demand, by the product of growth rate and tissue N. Uptake rates are reported as the slope of rate vs. concentration curves in each case. N uptake rates of U. curvata were inversely correlated with tissue N and affected only slightly by temperature. There was no correlation of N uptake rate with tissue N in C. decorticatum. N uptake rates of C. decorticatum were affected by temperature but to a lesser degree than were growth rates. Neither N addition per se nor light affected N uptake capacity of either species. The proximal mechanism for seaweeds accumulation of N at low light and temperatures may be that N uptake is less limited by light and temperature than is growth. This in turn may partially compensate for the effects of reduced light and temperature on growth by increasing pigment and enzyme levels. Perturbation uptake rates were higher than continuous mode or N demand rates in Ulva but not in Codium. N uptake rates of Ulva were higher than those of Codium, but N storage capacities were lower. These two observations suggest that Ulva experiences a fundamentally more variable N supply than does Codium. This is consistent with the clarification of Ulva as an ephemeral form and of Codium as persistent. A seaweed's functional form therefore appears to influence the spectrum of resource variability available to it as well as its ability to persist in the environment.     

EFFECTS OF NITROGEN AND PHOSPHORUS ON THE ENDOSYMBIONT LOAD OF RHOPALODIA GIBBA AND EPITHEMIA TURGIDA (BACILLARIOPHYCEAE)1

Diatoms of the family Epithemiaceae possess a unicellular nitrogen-fixing cyanobacterial endosymbiont. We investigated the potential of extracellular nitrogen and phosphorus concentrations to affect the endosymbiont load of Rhopalodia gibba O. Müll, and Epithemia turgida Ehr. in field and culture populations. In a growth chamber experiment, monoclonal cultures of R. gibba were exposed to three levels of nitrate-nitrogen. Nutrient-diffusing substrates were used in a lake environment to create nine microhabitats of varying nitrogen and phosphorus ratios for natural populations of R. gibba and E. turgida. The number of endosymbionts per diatom increased as ambient nitrogen became limiting; mean endosymbiont volume increased as nitrogen increased. The mean endosymbiont surface area: volume ratio decreased with increasing nitrogen. Total endosymbiont volume per diatom (the product of the number of endosymbionts per diatom and their individual biovolumes) did not have a simple response to increasing nitrogen. Phosphorus limitation uncoupled the relationship between endosymbiont load and nitrogen. We suspect that flexibility of the endosymbiont load can reduce the metabolic cost to the diatom if the endosymbionts are dependent on the diatom for a resource.     

PRIMARY ADHESION OF ENTEROMORPHA (CHLOROPHYTA,ULVALES) PROPAGULES: QUANTITATIVE SETTLEMENT STUDIES AND VIDEO MICROSCOPY1

Quantitative methods and associated kinetic analyses have been used for the first time to study detailed aspects of the settlement and adhesion of various types of Enteromorpha popagule. Time course experiments showed that quadri and biflagellate zoospores and zygotes adhered rapidly, but a proportion within any one population appeared to be incompetent at adhering to the substratum. Kinetic (Scatchard) analysis of adhesion experiments performed at a range of zoospore concentrations revealed density-dependent effects not previously reported, with positive cooperativity at low spore densities and negative cooperativity at high spore densities. High-resolution video microscopy was used for the first time to reveal details of the various stages in the settlement and adhesion of zoospores and zygotes. Novel observations were made of an initial, temporary phase of attachment via the apical papilla, followed by a permanent phase of commitment, characterized by discharge of adhesive-containing cytoplasmic vesicles, as the cell contracted against the surface, and adsorption of flagella. The phase of commitment was followed by exploitation of the surface through amoeboid-like movements at the interface. Gregarious settlement behavior was frequently observed leading to the formation of rafts of cells. The possible mechanisms and significance of density-dependent spore adhesion are discussed.     

PHOSPHORUS AND NITROGEN NUTRITION IN CHONDRUS CRISPUS (RHODOPHYTA): EFFECTS ON TOTAL PHOSPHORUS AND NITROGEN CONTENT,CARRAGEENAN PRODUCTION,AND PHOTOSYNTHETIC PIGMENTS AND METABOLISM1

The existence of a phenomenon in phosphorus (P) nutrition comparable to the “Neish effect” in nitrogen (N) nutrition (an inverse relation between seawater N enrichment and carrageenan content) was investigated in the temperate red alga Chondrus crispus Stackhouse. Plants were preconditioned for 17 d and then cultured under varying enrichments of P (0, 3, 6, 10, 15 μM P·wk^?1) and a constant N enrichment (53.5 μM N·wk^?1) for 5 wk. Tissue total P, tissue total N, and carrageenan contents were then determined. Identical experiments were performed using C. crispus collected during the fall, winter, spring, and summer seasons. The procedure was repeated using material collected during the following fall season and cultured under constant P (6 μM P·wk^?1) and varying N enrichments (0, 3, 6, 10, 25 μM N·wk^?1). In the fall (P) experiment, carrageenan content was the highest [53.1 ± 0.3% DW (dry weight)], and tissue total P content was the lowest (1.71 ± 0.27 mg P·g DW^?1) in plants that received no P enrichment. Carrageenan content was stable (46.1 ± 1.8% DW) for plants given enrichments of 3 μM P·wk^?1 and greater. Thus, a decrease in carrageenan content, concomitant with an increase in tissue total P content, was observed, but only at tissue total P levels below 2 mg P·g DW^?1. As these levels were always higher than 2 mg P·g DW^?1 in the winter, spring, and summer experiments, carrageenan content remained constant within each season at 46.2 ± 1.3, 43.1 m 0.7, and 44.5 ± 0.6% DW, respectively. Nitrogen enrichment of plants collected in the fall did not affect carrageenan content, which was stable at 49.3 ± 0.9% DW. When these plants were compared with those of the previous fall experiment (6 μM P·wk^?1 and 53.5 μM N·wk^?1), a slight increase in carrageenan content was noted. Thus, at sufficiently high concentration, N also decreased carrageenan content in C. crispus. Phosphorus nutrition had no significant effect on photosynthesis versus irradiance parameters (P_max, α, R_d, I_c, and I_k), the contents of the photosynthetic pigments chlorophyll-a, phycoerythrin (PE), phycocyanin (PC), and allophycocyanin (APC), and the ratios PE:APC and PC:APC. In contrast, N nutrition affected both P_maxand the photosynthetic pigment contents. The data indicate that N limitation reduces the number of phycobilisomes but not their size. The greater reduction in phycobiliprotein than chlorophyll-acontent corroborates the natural bleaching phenomenon regularly observed in C. crispus populations during summer when N levels are generally low in seawater. These results suggest that C. crispus in the temperate waters of the Bay of Fundy may experience N limitation, but P limitation is unlikely.     

EFFECT OF CURRENT VELOCITY ON THE DETACHMENT OF THALLI OF ULVA LACTUCA (CHLOROPHYTA) IN A NEW ZEALAND ESTUARY

Experiments were undertaken in a recirculating flume to determine the relationships among water velocity, thallus area, drag, and the probability of thallus breakage or detachment in the foliose green alga Ulva lactuca L. In all specimens tested to breaking point, thalli detached from their bivalve substrates as a result of stipe breakage rather than in midthallus or by holdfast detachment. There was no relationship between thallus size and drag at which detachment occurred. Rather, the probability of detachment was normally distributed about a mean drag of 0. 70 N (95% confidence limits 0.55–0.85 N). Average breaking stress of stipes was 345 kN.m^-2 (95% cl 250–485 kN.m^-2). Similar results were obtained in field experiments where the horizontal force required to detach thalli was measured directly as 0.93 N (95% cl 0.69–1.15 N). Drag coefficients of plants were not constant with water velocity but increased up to 0.4 m.s^-1, declining exponentially at velocities above this. Empirical relationships were established between coefficient of drag and Reynold's number and, hence, among drag, thallus area and water velocity. These relationships permitted estimation of mean water velocity at which plants of a given area would detach .     

GAMETOGENESIS AND GAMETE RELEASE OF ULVA MUTABILIS AND ULVA LACTUCA (CHLOROPHYTA): REGULATORY EFFECTS AND CHEMICAL CHARACTERIZATION OF THE “SWARMING INHIBITOR”1

Gametophytes of Ulva mutabilis Føyn and Ulva lactuca L. were artificially induced to form gametangia by removal of sporulation inhibitors. After this treatment, U. mutabilis gametes were ready for swarming on the third morning after induction, while U. lactuca gametangia needed 1–2 d longer for maturation. Release of gametes of U. lactuca was dependent solely upon exposure to the first light in the morning. Gametangia of U. mutabilis, however, also required sufficient dilution of the swarming inhibitor (SWI). SWI was excreted transiently by both Ulva species early during gametogenesis. While the SWI concentration in U. mutabilis medium remained above the inhibitory concentration until the gametangia were mature, the concentration of U. lactuca‐SWI dropped rapidly below this level. In the presence of sufficient SWI, mature gametes of U. mutabilis remained motionless within the gametangia despite light and open exit pores. However, using SEM, an additional seal was detected within these pores, which probably prevented premature swarming until dilution of SWI and exposure to light. Observations by time lapse microscopy and experiments with the myosin kinase inhibitor BDM suggest that the gametes may be either extruded by the gametangium or leave the exit pore by active gliding motion, driven by a myosin‐like motor protein. The SWIs were purified from both Ulva species, and mass spectral analysis showed their molecular masses (292 Da) were identical.     

TRANSIENT AMMONIUM UPTAKE IN THE MACROALGA ULVA LACTUCA (CHLOROPHYTA): NATURE,REGULATION, AND THE CONSEQUENCES FOR CHOICE OF MEASURING TECHNIQUE1

The nature of transient ammonium uptake by the macroalga Ulva lactuca L. was studied from the depletion of ammonium after single additions of ammonium to batch cultures. The experiments were carried out by the application of two different experimental setups: the “multiple flask” and the “perturbation” techniques. Uptake rate was nonlinear with time, and three distinct, succeeding phases of uptake were identified: 1) “surge” uptake, i.e. transiently enhanced uptake that lasted for a few hours only, followed by 2) “internally” controlled uptake, a relatively constant phase occurring at high substrate concentrations, and finally 3) the “externally” controlled uptake phase, which was substrate-dependent and occurred at low substrate concentrations. Surge uptake occurred over a broad range of substrate concentrations but was concentration-dependent and, so, equalled externally controlled uptake rates at substrate concentrations below 3–10 μM. The transient nature of ammonium uptake rate seemed related to rapid changes in small intracellular pools of inorganic nitrogen or amino acids rather than to changes in total N content of the algae. The transient nature of ammonium uptake has important implications for the measurement of uptake rates when either of the two standard methods, the multiple flask and the perturbation technique, are used, and I recommend that a combination of the two methods be used for future uptake experiments.     

EFFECT OF NITROGEN SOURCE ON PLATYMONAS (CHLOROPHYTA) CELL COMPOSITION AND AMINO ACID UPTAKE RATES1

The effect of nitrogen source (nitrate, ammonia and/or amino acids) on cell composition and amino acid uptake rates was examined. Substantial levels of free amino acids accumulated intracellularly with all nitrogen sources used. Ammonia accumulated only when provided in the medium. The presence of ammonia in the medium decreased the intracellular accumulation of free amino acids, especially arginine. Amino acid uptake rates were suppressed by the presence of excess nitrogen, especially ammonia. However, the suppression of uptake did not show any particular relation to the nitrogenous cell composition.     

MOLECULAR ASSESSMENT OF ULVA SPP. (ULVOPHYCEAE,CHLOROPHYTA) IN THE HAWAIIAN ISLANDS1

Sequences of the nuclear internal transcribed spacer 1 (ITS1) region and the chloroplast rbcL gene were obtained from 86 specimens of Ulva (including “Enteromorpha”) from five of the main Hawaiian Islands. These 86 specimens were divided into 11 operational taxonomic units (OTUs) based on analyses of primary sequence data and comparisons of ITS1 secondary structure. Of the 11 OTUs, six have not previously been reported from anywhere in the world. Only three represented exact sequence matches to named species (Ulva lactuca L., syn. U. fasciata Delile; U. ohnoi Hiraoka et Shimada); two others represented exact sequence matches to unnamed species from Japan and New Zealand. Of the 12 species names currently in use for Hawaiian Ulva, only one, U. lactuca (as U. fasciata), was substantiated. General morphology of the specimens did not always correspond with molecular OTUs; for example, reticulate thallus morphology, previously considered diagnostic for the species U. reticulata Forssk., was expressed in thalli assigned to U. ohnoi and to one of the novel OTUs. This finding confirms a number of recent studies and provides further support for a molecular species concept for Ulva. These results suggest that Ulva populations in tropical and subtropical regions consist of species that are largely unique to these areas, for which the application of names based on types from temperate and boreal European and North American waters is inappropriate. Ulva ohnoi, a “green tide” species, is reported from Hawaii for the first time.